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Sensitivity of larvae and adult and the immunologic characteristics of Litopenaeus vannamei under the acute hypoxia.

1. Introduction

In recent years, because of human activities and the changes of natural environment, the severity, frequency of occurrence, and duration of hypoxia are increasing, resulting in high mortality of valuable living resources [1, 2]. Hypoxia can make coastal benthic ecosystems generate so-called dead zones, and it was reported that more than four hundred ecosystems generated dead zones [3, 4]. Hypoxia may occur as a result of single factor or combined action of several factors such as eutrophication, stratification of the water column, freshwater runoff, weather, and biological processes. However, water eutrophication caused by human activities is the main reason of deterioration and increase in hypoxia zones [5-7]. White shrimp (Litopenaeus vannamei) is a very commercially important species in the world. However, hypoxia has seriously affected the growth and development and even resulted in mortality of shrimp [8].

Previous researches had shown that juvenile Metapenaeus ensis had 25% mortality at dissolved oxygen (DO) levels between 0.35 and 0.60 ppm, 8.3% mortality at DO levels between 0.60 and 0.85 mg [L.sup.-1], and 0% mortality at DO levels between 1.0 and 1.36 mg [L.sup.-1] in 24 h lab experiment [9]. The 24 h median lethal concentration ([LC.sub.50]) values for 3- and 10-day-old mysids were 1.51 mg [L.sup.-1] and 1.56 mg [L.sup.-1], respectively, and 24 h and 48 h [LC.sub.50] for pink shrimp (Penaeus duorarum) whose mean total length was 90.8 mm were 1.36 and 1.46 mg [L.sup.-1], respectively. So these results show that larvae are more sensitive than adult to hypoxia and that is similar to red drum (Sciaenops ocellatus) 10]. The majority of mortality owing to hypoxia happened in the first four hours of hypoxia and the test duration of more than 24 h had little effect on medium lethal concentration, and most of the [LC.sub.50] values for fish at 2-4 days fell within 0.1 mg [L.sup.-1] compared with that obtained from 24 h [11].

Marine invertebrates, including mussels (e.g., Mytilusgalloprovincialis) [12, 13], clams (e.g., Ruditapes philippinarum) [14-17], oysters (e.g., Crassostrea gigas) 18], and shrimps, are widely studied in immunology and environmental science [19-23]. Because of the lack of the acquired immune system, marine invertebrates, such as shrimp, just rely on innate immune system to resist environmental stress [24, 25]. Humoral immunity includes antimicrobial peptides, clotting cascade, and prophenoloxidase, and the cellular defenses of shrimp depend on hemocytes that have many functions such as production of antimicrobial compounds, apoptosis, nodule formation, encapsulation of pathogens, cell adhesion, wound healing, coagulation, and phagocytosis [26, 27]. Phagocytosis and apoptosis play an important role in shrimp response to virus infection [28]. Hypoxia can evidently decrease the total hemocyte counts (THC), bacteriolytic activity, antibacterial activity, phenoloxidase activity, and phagocytic activity of prawn [29, 30]. Therefore, hypoxia can increase shrimp sensibility to pathogens and decrease defense capability to diseases.

Hemocyanin is an important respiratory protein and a major plasmatic protein in crustaceans and plays an important role in binding and transporting oxygen and C[O.sub.2] [31]. Moreover, the cleaved fragments of hemocyanin have antibacterial activity and may improve immunity of shrimp [32]. Under the condition of hypoxia, shrimp can increase the hemocyanin concentration (HC) and the affinity of hemocyanin for oxygen to improve the tolerance to hypoxia [33]. The concentration of copper ion reduces in hepatopancreas; however, the HC and copper ion increase in the hemolymph under hypoxia. These may suggest that shrimp can transport copper ion to compound hemocyanin to enhance the capability of carrying oxygen. Hypoxia also can make the gene expression of hemocyanin increase [34].

Most of previous studies focused on measuring the [LC.sub.50] values in the special life stage and HC and THC in the condition of hypoxia, but there was very little information about the [LC.sub.50] values in the whole life stages in the condition of hypoxia and the changes of HC and THC in the phase of reoxygenation. So, we performed studies to determine the [LC.sub.50] values in whole life stages and investigated the changes of HC and THC in the condition of hypoxia and reoxygenation. These results can better understand the physiological mechanism of shrimp in the condition of hypoxia. Furthermore, it can provide fundamental data for shrimp farming and seedling.

2. Materials and Methods

2.1. Animals. Litopenaeus vannamei were obtained from a farm at Wenchang of Hainan province in China. The larvae of shrimp were directly collected from nursery pond, and the adult shrimps were acclimated in tank containing seawater (the salinity, temperature, and pH are displayed in Table 1) for three days before experiment. Half of the seawater in tank was replaced once daily and the shrimps were fed with formulated shrimp diet twice daily during the acclimation period. The shrimps were not fed during the experiment period.

2.2. Experimental Design. The first experiment was conducted to determine the [LC.sub.50] values at different life stages of Litopenaeus vannamei. The larvae shrimps were cultivated in a 5 L beaker, and six dissolved oxygen levels (0.43, 0.85, 1.70, 2.56, 3.40, and 5.00 mg [L.sup.-1]) were established. The adult shrimps were cultivated in tanks (75 L) and five dissolved oxygen levels (0.3, 0.5,1.0, 2.0, and 6.0 mg [L.sup.-1]) were established. Each dissolved oxygen level was conducted in triplicate, and 50 shrimps were contained in each replicate. Test conditions were presented in Table 1. After 12 hours, the numbers of deaths and survivals were counted, respectively.

The second experiment was conducted to determine the changes of HC and THC in the phase of hypoxia and reoxygenation. The average body lengths of shrimp were 125 [+ or -] 0.5 mm and the animals were acclimated in tank containing seawater (salinity 31, pH 7.89, and temperature 27 [+ or -] 1[degrees]C) for three days before use. Three dissolved oxygen levels (1.0, 3.0, and 7.0 mg [L.sup.-1]) were established and each level was conducted in triplicate. Each test tank (75 L) contained 30 shrimps.

The desired dissolved oxygen levels were established by bubbling nitrogen gas and air into the seawater, and the dissolved oxygen levels were monitored with a DO meter (HI 9146, HANNA) every half an hour. The pH and salinity values were measured by pH meter (HI 8424, HANNA) and salinity meter (RSH-28), respectively.

2.3. Hemolymph Collection and Total Hemocyte Counts (THC). Hemolymph was collected randomly from each replicate at 0, 3, 6, 12, 18, and 24 h during the period of hypoxia and reoxygenation. Hemolymph was withdrawn individually from the cardiocoelom of shrimp with a 1.0 mL syringe filled with an equal volume of anticoagulant solution (30 mM trisodium citrate, 0.34 M sodium chloride, 10 mM EDTA, and 0.115 M glucose pH 755) and stored, respectively, in 1.5 mL Eppendorf centrifuge tubes. 30 [micro]L of anticoagulant hemolymph was added immediately into the blood counting chamber by micropipette. Then hemocytes were observed under the optical microscope (Olympus) and THC were recorded by a cell counter. Another anticoagulant hemolymph was stored at -20[degrees]C for HC assay.

Anticoagulant hemolymph was centrifuged at 800 g for 10 min under 4[degrees]C for HC assay. Then 100 [micro]L supernatant fraction was moved individually into 5 mL centrifuge tubes and diluted 1: 30 with Tris-Ca buffer (50 mM Tris, 10 mM Ca[Cl.sub.2], and pH = 8.0). The OD values of the diluted plasma were measured at 335 nm using a UV spectrophotometer (1cm path length) (PerkinElmer Lambda 25), and hemocyanin concentration (unit: mg [mL.sup.-1]) was calculated using the following formula: E335 nm (mg [mL.sup.-1]) = 2.3 x O[D.sub.335 nm] (E stands for HC; 2.3 is the extinction coefficient of hemocyanin for mg [mL.sup.-1]) 35, 36].

2.4. Statistical Analyses. All data were tackled using SPSS19.0. Suppose that P < 0.05 was the significant level. All data in the present study were shown as mean [+ or -] SD (n = 3). All figures were made by Origin 8.0.

3. Results

3.1. The Estimated 12 h [LC.sub.50], 90% Lethal Concentration ([LC.sub.90]), and 10% Lethal Concentration ([LC.sub.10]) for Dissolved Oxygen at Different Life Stages of Shrimp Litopenaeus vannamei. The estimated [LC.sub.50], [LC.sub.90], and [LC.sub.10] were shown in Table 2. The results indicated that the life stages had a significant influence on [LC.sub.90], [LC.sub.50], and [LC.sub.10] values (P < 0.01). The highest [LC.sub.50] value was 2.113 mg [L.sup.-1] in the phase of mysis III, and the lowest [LC.sub.50] value was 0.535 mg [L.sup.-1] in the phase of 6 cm. The results showed that larvae were more sensitive to hypoxia than adult and the changed trend of [LC.sub.50], [LC.sub.90], and [LC.sub.10] values was similar at whole life stages of shrimp. The range of [LC.sub.50] in the phase of larvae (1.039 mg [L.sup.-1]) was larger than adult shrimp (0.116 mg [L.sup.-1]).

3.2. Effects of Hypoxia and Reoxygenation on THC and HC in Hemolymph ofL. vannamei. Figures 1 and 2 showed that hypoxia had an obvious influence on HC and THC (P < 0.05). HC and THC had no significant difference in the control (7 mg [L.sup.-1]) (P > 0.05).

Figure 1 indicated that HC was significantly increased under the condition of hypoxia and decreased after reoxygenation, respectively. The HC returned to normal level after reoxygenation for 24 h.

Figure 2 demonstrated that THC obviously decreased in the phase of hypoxia and increased after reoxygenation. The result showed that the more serious and longer the duration of hypoxia, the more the reduction of THC. The THC of treatment group fell to the lowest level after reoxygenation for 3 h, and it did not return to normal level after reoxygenation for 24 h. The THC in the treatment groups had significant difference (P < 0.05) compared with control groups.

4. Discussion and Conclusion

The estimated [LC.sub.50] values are different at different life stages of white shrimp, and the larvae stage is more sensitive than adult stage. The range of [LC.sub.50] in the phase of larvae (1.039 mg[L.sup.-1]) is larger than adult shrimp (0.116 mg [L.sup.-1]). The reasons may be that the growth and development of adult shrimp are more perfect and they can more efficiently resist environmental stresses.

Previous researches indicated that the 24 h [LC.sub.50] values for 3- and 10-day-old mysids were 1.51 and 1.56 ppm, respectively; these are similar to the 12 h [LC.sub.50] in this work. The 24 h and 48 h [LC.sub.50] of pink shrimp were 1.36 and 1.46 mg [L.sup.-1], respectively [10], and the average total length of pink shrimp was about 90.8 mm; the values are bigger than the 12 h [LC.sub.50] in this work. Majority of mortality owing to hypoxia happened in the first four hours of hypoxia and the test duration of more than 24 h had little effect on [LC.sub.50], and most of the [LC.sub.50] values for fish at 2-4 days fell within 0.1 ppm compared with that obtained from 24 h [11]. This indicates that shrimps may have adapted to hypoxic environment by adjusting their behaviors [37], physiological and biochemical functions [31, 32], respiratory metabolism [38], and even gene expression [39] after a period of treatment.

Mysis III is the most sensitive stage to hypoxia in whole life and the key phase in the process of shrimp culture. The [LC.sub.50] values in the present study are little different from the values of published literature [40]. It may be due to the different sensitivities at different life stages, test conditions, and species of shrimp.

With the increasing experimental time, the change tendency of HC and THC in the present study was similar to published literature [31]. Hemocyanin is the main plasmatic protein and has ability to bind and transport oxygen and metabolically produced C[O.sub.2] in crustaceans [41]. In the condition of hypoxia, the increase in HC can improve the ability of shrimp to gain more oxygen to deal with hypoxic stress. After reoxygenation, the decrease in HC indicated that shrimp no longer needed so much hemocyanin to gain oxygen. Published literature reported that the majority of mortality owing to hypoxia happened in the first four hours of exposure and the test duration of more than 24 h had little effect on lethal concentration [10]. In the first three hours of hypoxia, the increasing rate of HC and the decreasing rate of THC are larger than other hypoxia periods; it suggests that shrimp can rapidly activate a complicated system to respond to the hypoxia stress.

Shrimp only has innate immune system including hemocyte and diverse active factors existing in hemocyte or released to hemolymph from the hemocyte, so hemocyte plays an important role in immune defense. The results of the present and previous studies showed that THC, bacteriolytic activity, antibacterial activity, phagocytic activity, and phenoloxidase activity decreased significantly and the sensibility to pathogens increased in the condition of hypoxia [42]. Therefore, we can deduce that hypoxia can reduce the immune defense of shrimp. These results will contribute to the cultivation and seedling of shrimp and also be helpful to better understand the stress mechanism of hypoxia, but there is still much work which needs to be done; for example, we need to use omic techniques to explore the molecular mechanism of hypoxia stress [43] and need to develop some specific biomarkers to monitor the hypoxia stress; there has been useful information in other invertebrates, such as Mytilus galloprovincialis [44-46] and Venerupis philippinarum [47].

http://dx.doi.org/10.1155/2014/574534

Disclosure

Yuhu Li is the first coauthor.

Conflict of Interests

The authors declare that there is no conflict of interests regarding the publication of this paper.

Acknowledgments

This work was supported by the China Postdoctoral Science Foundation Funded Project (2013M530332), the Specialized Research Fund for the Midwest Programme of Hainan University (ZXBJH-XK002), the Specialized Research Fund for the Doctoral Programme of Higher Education of China (20114601120001), and the Hainan Province Special Fund of the Integration of Industrialization, Teaching and Research (CXY20130054).

References

[1] E. D. Goldberg, "Emerging problems in the coastal zone for the twenty-first century," Marine Pollution Bulletin, vol. 31, no. 4-12, pp. 152-158, 1995.

[2] R. S. S. Wu, "Eutrophication, water borne pathogens and xenobiotic compounds: environmental risks and challenges," Marine Pollution Bulletin, vol. 39, no. 1-12, pp. 11-22, 1999.

[3] N. N. Rabalais, R. E. Turner, and D. Scavia, "Beyond Science into Policy: Gulf of Mexico Hypoxia and the Mississippi River Nutrient policy development for the Mississippi River watershed reflects the accumulated scientific evidence that the increase in nitrogen loading is the primary factor in the worsening of hypoxia in the northern Gulf of Mexico," BioScience, vol. 52, no. 2, pp. 129-142, 2002.

[4] R. J. Diaz and R. Rosenberg, "Spreading dead zones and consequences for marine ecosystems," Science, vol. 321, no. 5891, pp. 926-929, 2008.

[5] N. N. Rabalais, R. E. Turner, B. K. Sen Gupta, D. F. Boesch, P. Chapman, and M. C. Murrell, "Hypoxia in the northern Gulf of Mexico: does the science support the plan to reduce, mitigate, and control hypoxia?" Estuaries and Coasts, vol. 30, no. 5, pp. 753-772, 2007.

[6] L. Stramma, G. C. Johnson, J. Sprintall, and V. Mohrholz, "Expanding oxygen-minimum zones in the tropical oceans," Science, vol. 320, no. 5876, pp. 655-658, 2008.

[7] R. McAllen, J. Davenport, K. Bredendieck, and D. Dunne, "Seasonal structuring of a benthic community exposed to regular hypoxic events," Journal of Experimental Marine Biology and Ecology, vol. 368, no. 1, pp. 67-74, 2009.

[8] A. Haselmair, M. Stachowitsch, M. Zuschin, and B. Riedel, "Behaviour and mortality of benthic crustaceans in response to experimentally induced hypoxia and anoxia in situ," Marine Ecology Progress Series, vol. 414, pp. 195-208, 2010.

[9] S. Yamochi, H. Ariyama, and M. Sano, "Occurrence and hypoxic tolerance of the juvenile Metapenaeus ensis at the mouth of the Yodo River, Osaka," Fisheries Science, vol. 61, no. 3, pp. 391-395, 1995.

[10] L. R. Goodman and J. G. Campbell, "Lethal levels of hypoxia for gulf coast estuarine animals," Marine Biology, vol. 152, no. 1, pp. 37-42, 2007.

[11] D. C. Miller, S. L. Poucher, and L. Coiro, "Determination of lethal dissolved oxygen levels for selected marine and estuarine fishes, crustaceans, and a bivalve," Marine Biology, vol. 140, no. 2, pp. 287-296, 2002.

[12] C. Ji, H. Wu, L. Wei, J. Zhao, Q. Wang, and H. Lu, "Responses of Mytilus galloprovincialis to bacterial challenges by metabolomics and proteomics," Fish and Shellfish Immunology, vol. 35, no. 2, pp. 489-498, 2013.

[13] X. Liu, C. Ji, J. Zhao, Q. Wang, F. Li, and H. Wu, "Metabolic profiling of the tissue-specific responses in mussel Mytilus galloprovincialis towards Vibrio harveyi challenge," Fish & Shellfish Immunology, vol. 39, no. 2, pp. 372-377, 2014.

[14] X. Liu, J. Zhao, H. Wu, and Q. Wang, "Metabolomic analysis revealed the differential responses in two pedigrees of clam Ruditapes philippinarum towards Vibrio harveyi challenge," Fish and Shellfish Immunology, vol. 35, no. 6, pp. 1969-1975, 2013.

[15] X. Liu, C. Ji, J. Zhao, and H. Wu, "Differential metabolic responses of clam Ruditapes philippinarum to Vibrio anguillarum and Vibrio splendidus challenges," Fish and Shellfish Immunology, vol. 35, no. 6, pp. 2001-2007, 2013.

[16] C. Li, H. Sun, A. Chen et al., "Identification and characterization of an intracellular Cu, Zn-superoxide dismutase (icCu/ZnSOD) gene from clam Venerupis philippinarum," Fish and Shellfish Immunology, vol. 28, no. 3, pp. 499-503, 2010.

[17] J. Zhao, L. Qiu, X. Ning, A. Chen, H. Wu, and C. Li, "Cloning and characterization of an invertebrate type lysozyme from Venerupis philippinarum," Comparative Biochemistry and Physiology B: Biochemistry and Molecular Biology, vol. 156, no. 1, pp. 56-60, 2010.

[18] M. Cong, J. Lu, H. Wu, and J. Zhao, "Effect of cadmium on the defense response of Pacific oyster Crassostrea gigas to Listonella anguillarum challenge," Chinese Journal of Oceanology and Limnology, vol. 31, no. 5, pp. 1002-1009, 2013.

[19] L. You, X. Ning, F. Liu, J. Zhao, Q. Wang, and H. Wu, "The response profiles of HSPA12A andTCTP from Mytilus galloprovincialis to pathogen and cadmium challenge," Fish and Shellfish Immunology, vol. 35, no. 2, pp. 343-350, 2013.

[20] L. Zhang, X. Liu, L. You et al., "Metabolic responses in gills of Manila clam Ruditapes philippinarum exposed to copper using NMR-based metabolomics," Marine Environmental Research, vol. 72, no. 1-2, pp. 33-39, 2011.

[21] X. Liu, L. Zhang, L. You et al., "Toxicological responses to acute mercury exposure for three species of Manila clam Ruditapes philippinarum by NMR-based metabolomics," Environmental Toxicology and Pharmacology, vol. 31, no. 2, pp. 323-332, 2011.

[22] L. Zhang, X. Liu, L. You et al., "Benzo(a)pyrene-induced metabolic responses in Manila clam Ruditapes philippinarum by proton nuclear magnetic resonance ([sup.1]H NMR) based metabolomics," Environmental Toxicology and Pharmacology, vol. 32, no. 2, pp. 218-225, 2011.

[23] L. Zhang, L. Qiu, H. Wu et al., "Expression profiles of seven glutathione S-transferase (GST) genes from Venerupis philippinarum exposed to heavy metals and benzo[a]pyrene," Comparative Biochemistry and Physiology C: Toxicology and Pharmacology, vol. 155, no. 3, pp. 517-527, 2012.

[24] E. Bachere, Y. Gueguen, M. Gonzalez, J. de Lorgeril, J. Garnier, and B. Romestand, "Insights into the anti-microbial defense of marine invertebrates: the penaeid shrimps and the oyster Crassostrea gigas" Immunological Reviews, vol. 198, no. 1, pp. 149-168, 2004.

[25] W. Cheng and J.-C. Chen, "Effects of pH, temperature and salinity on immune parameters of the freshwater prawn Macrobrachium rosenbergii" Fish and Shellfish Immunology, vol. 10, no. 4, pp. 387-391, 2000.

[26] T. J. Little, D. Hultmark, and A. F. Read, "Invertebrate immunity and the limits of mechanistic immunology," Nature Immunology, vol. 6, no. 7, pp. 651-654, 2005.

[27] F. Li and J. Xiang, "Recent advances in researches on the innate immunity of shrimp in China," Developmental and Comparative Immunology, vol. 39, no. 1-2, pp. 11-26, 2013.

[28] L. Wang, B. Zhi, W. Wu, and X. Zhang, "Requirement for shrimp caspase in apoptosis against virus infection," Developmental and Comparative Immunology, vol. 32, no. 6, pp. 706-715, 2008.

[29] L.-X. Jiang and L.-Q. Pan, "Effect of dissolved oxygen on immune parameters of the white shrimp Litopenaeus vannamei" Fish and Shellfish Immunology, vol. 18, no. 2, pp. 185-188, 2005.

[30] F. Hu, L. Pan, and F. Jing, "Effects of hypoxia on dopamine concentration and the immune response of white shrimp (Litopenaeus vannamei)," Journal of Ocean University of China, vol. 8, no. 1, pp. 77-82, 2009.

[31] W. Cheng, C.-H. Liu, and C.-M. Kuo, "Effects of dissolved oxygen on hemolymph parameters of freshwater giant prawn, Macrobrachium rosenbergii (deMan)" Aquaculture, vol. 220, no. 1-4, pp. 843-856, 2003.

[32] D. Destoumieux-Garzon, D. Saulnier, J. Garnier, C. Jouffrey, P. Bulet, and E. Bachere, "Crustacean immunity: Antifungal peptides are generated from the C terminus of shrimp hemocyanin in response to microbial challenge," The Journal of Biological Chemistry, vol. 276, no. 50, pp. 47070-47077, 2001.

[33] J.-C. Chen and T.-T. Kou, "Hemolymph acid-base balance, oxyhemocyanin, and protein levels of Macrobrachium rosenbergii at different concentrations of dissolved oxygen," Journal ofCrustacean Biology, vol. 18, no. 3, pp. 437-441, 1998.

[34] M. Brouwer, N. J. Brown-Peterson, P. Larkin et al., "Molecular and whole animal responses of grass shrimp, Palaemonetes pugio, exposed to chronic hypoxia," Journal of Experimental Marine Biology and Ecology, vol. 341, no. 1, pp. 16-31, 2007.

[35] L. Yang and L. Pan, "Effects of phosphatidyl serine on immune response in the shrimp Litopenaeus vannamei" Central European Journal of Biology, vol. 8, no. 11, pp. 1135-1144, 2013.

[36] K. W. Nickerson and K. E. van Holde, "A comparison of molluscan and arthropod hemocyanin-I. Circular dichroism and absorption spectra," Comparative Biochemistry and Physiology Part B: Biochemistry and, vol. 39, no. 4, pp. 855-872, 1971.

[37] R. J. A. Atkinson and A. C. Taylor, "Aspects of the physiology, biology and ecology of thalassinidean shrimps in relation to their burrow environment," Oceanography and Marine Biology, vol. 43, pp. 173-210, 2005.

[38] I. S. Racotta, E. Palacios, and L. Mondez, "Metabolic responses to short and long-term exposure to hypoxia in white shrimp (Penaeus Vannamei)" Marine and Freshwater Behaviour and Physiology, vol. 35, no. 4, pp. 269-275, 2002.

[39] K. Kodama, M. S. Rahman, T. Horiguchi, and P. Thomas, "Assessment of hypoxiainducible factor-1a mRNA expression in mantis shrimp as a biomarker of environmental hypoxia exposure," Biology Letters, vol. 8, no. 2, pp. 278-281, 2012.

[40] R. S. S. Wu, P K. S. Lam, and K. L. Wan, "Tolerance to, and avoidance of, hypoxia by the penaeid shrimp (Metapenaeus ensis)," Environmental Pollution, vol. 118, no. 3, pp. 351-355, 2002.

[41] A. Tyler and C. B. Metz, "Natural heteroagglutinins in the serum of the spiny lobster, Panulirus interruptus. I. Taxonomic range of activity, electrophoretic and immunizing properties," Journal of Experimental Zoology, vol. 100, no. 3, pp. 387-406, 1945.

[42] W. Cheng, C.-H. Liu, J.-P Hsu, and J.-C. Chen, "Effect of hypoxia on the immune response of giant freshwater prawn Macrobrachium rosenbergii and its susceptibility to pathogen Enterococcus" Fish and Shellfish Immunology, vol. 13, no. 5, pp. 351-365, 2002.

[43] H. Wu, C. Ji, L. Wei, J. Zhao, and H. Lu, "Proteomic and metabolomic responses in hepatopancreas of Mytilus galloprovincialis challenged by Micrococcus luteus and Vibrio anguillarum" Journal of Proteomics, vol. 94, pp. 54-67, 2013.

[44] Q. Wang, C. Wang, C. Mu, H. Wu, L. Zhang, and J. Zhao, "A Novel C-Type lysozyme from Mytilus galloprovincialis: insight into innate immunity and molecular evolution of invertebrate C-type lysozymes," PLoS ONE, vol. 8, no. 6, Article ID e67469, 2013.

[45] Q. Wang, Z. Yuan, H. Wu, F. Liu, and J. Zhao, "Molecular characterization of a manganese superoxide dismutase and copper/zinc superoxide dismutase from the mussel Mytilus galloprovincialis" Fish and Shellfish Immunology, vol. 34, no. 5, pp. 1345-1351, 2013.

[46] Q. Wang, L. Zhang, J. Zhao, L. You, and H. Wu, "Two goose-type lysozymes in Mytilus galloprovincialis: possible function diversification and adaptive evolution," PLoS ONE, vol. 7, no. 9, Article ID e45148, 2012.

[47] L. Zhang, X. Liu, L. Chen et al., "Transcriptional regulation of selenium-dependent glutathione peroxidase from Venerupis philippinarum in response to pathogen and contaminants challenge," Fish and Shellfish Immunology, vol. 31, no. 6, pp. 831-837, 2011.

Hailong Zhou, (1,2) Yuhu Li, (2) Lin Wei, (2) Zhihuai Zhang, (1) Hao Huang, (3) Xiaoping Diao, (2) and Jianhai Xiang (1)

(1) Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China

(2) College of Agriculture, Hainan University, Haikou 570228, China

(3) Hainan Guangtai Marine Animal Breeding Limited Company, Wenchang 571328, China

Correspondence should be addressed to Xiaoping Diao; diaoxip@hainu.edu.cn and Jianhai Xiang; jhxiang@qdio.ac.cn

Received 9 October 2014; Accepted 12 November 2014; Published 25 November 2014

Academic Editor: Yongjun Gao

TABLE 1: Experiment conditions to determine the [LC.sub.50] for
dissolved oxygen of white shrimp Litopenaeus vannamei.

                                                          Salinity
                   Mean total      Temperature             ([per
Life stage        length (mm)     ([degrees]C)     pH    thousand])

Zygote                 --         29 [+ or -] 1   7.96       30
Nauplius               --         30 [+ or -] 1   7.98       30
Zoea I                 --         30 [+ or -] 1   7.96       30
Zoea II                --         29 [+ or -] 1   7.98       31
Zoea III               --         29 [+ or -] 1   7.95       31
Mysis I                --         30 [+ or -] 1   7.96       30
Mysis II               --         30 [+ or -] 1   7.91       30
Mysis III              --         30 [+ or -] 1   7.91       30
Postlarvae I           --         30 [+ or -] 1   7.93       30
Postlarvae II          --         31 [+ or -] 1   7.89       30
Postlarvae III         --         30 [+ or -] 1   7.92       30
Postlarvae IV          --         30 [+ or -] 1   7.88       30
Postlarvae V           --         29 [+ or -] 1   7.88       30
Postlarvae VI          --         29 [+ or -] 1   7.82       30
Adult            50 [+ or -] 2    29 [+ or -] 1   7.92       31
Adult            60 [+ or -] 4    28 [+ or -] 1   7.91       31
Adult            75 [+ or -] 4    29 [+ or -] 1   7.93       31
Adult            85 [+ or -] 5    26 [+ or -] 1   7.92       31
Adult            100 [+ or -] 5   23 [+ or -] 1   7.95       31
Adult            115 [+ or -] 3   22 [+ or -] 1   7.96       31
Adult            133 [+ or -] 5   25 [+ or -] 1   7.79       31

TABLE 2: Estimated 12 h [LC.sub.50], 95% confidence intervals (CI),
90% lethal concentration, and 10% lethal concentration of the test
white shrimp Litopenaeus vannamei.

                                              90% lethal
Life stage       [LC.sub.50]       CI        concentration

Zygote              1.288                        0.546
Nauplius            1.335      0.899-1.737       0.868
Zoea I              1.404      1.257-1.525       1.057
Zoea II             1.387      1.055-1.611       1.046
Zoea III            1.722      1.569-1.856       1.230
Mysis I             1.722      1.569-1.856       1.230
Mysis II            1.762      1.600-1.910       1.178
Mysis III           2.113      1.967-2.254       1.537
Postlarvae I        1.490      1.340-1.632       0.951
Postlarvae II       1.504      1.353-1.651       0.920
Postlarvae III      1.349      1.197-1.502       0.709
Postlarvae IV       1.074      0.954-1.197       0.607
Postlarvae V        1.135      1.006-1.266       0.620
Postlarvae VI       1.299      1.153-1.446       0.696
5 cm                0.577      0.463-0.775       0.377
6 cm                0.535      0.430-0.731       0.351
75 cm               0.625      0.404-1.477       0.429
8.5 cm              0.640      0.564-0.735       0.419
10 cm               0.593      0.537-0.762       0.463
11.5 cm             0.593      0.537-0.762       0.463
13.5 cm             0.651      0.582-0.740       0.482

                  10% lethal
Life stage       concentration

Zygote               3.042
Nauplius             2.054
Zoea I               1.866
Zoea II              1.838
Zoea III             2.413
Mysis I              2.413
Mysis II             2.637
Mysis III            2.907
Postlarvae I         2.335
Postlarvae II        2.460
Postlarvae III       2.564
Postlarvae IV        1.900
Postlarvae V         2.076
Postlarvae VI        2.424
5 cm                 0.883
6 cm                 0.816
75 cm                0.911
8.5 cm               0.977
10 cm                0.759
11.5 cm              0.759
13.5 cm              0.878

Note. The results were derived from the mean of repeated experiments
and the duration of hypoxia was 12 hours.
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Title Annotation:Research Article
Author:Zhou, Hailong; Li, Yuhu; Wei, Lin; Zhang, Zhihuai; Huang, Hao; Diao, Xiaoping; Xiang, Jianhai
Publication:Journal of Chemistry
Article Type:Report
Date:Jan 1, 2014
Words:4762
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